Clinical-scale Production of Osteoprogenitor Cells

描述（由申请人提供）：由于发病率和材料量有限，需要自体移植物的替代品进行骨修复。已知骨髓内的细胞具有骨再生能力，但它们的数量有限。通过使用离体扩增技术增加这些骨祖细胞群并将这些细胞与基质材料组合可显著增加新骨形成的量和速率，并取代对自体移植物的需求。I期研究确定了骨祖细胞扩增的扩增变量，并表明该过程可以扩大到自动化临床规模的细胞培养系统，AastromReplicell（r）细胞生产系统（ARS）。在II期研究的目标1中，将继续优化骨祖细胞扩张过程。优化将不仅通过体外和体内的生物学性能来衡量，而且还通过扩增过程的商业化容易程度来衡量。具体而言，将确定其他细胞群在骨祖细胞扩增和功能中的作用，将确定在培养物中包含外源性因子的最低要求，将确定接种密度和最大化补料方案以支持从小体积骨髓抽吸物扩增，将检查在无动物血清环境中扩增的潜力。在目的2中，将通过集落测定和管形成测定在体外测量这些相同细胞产物的血管生成潜力，因为血管形成对于最佳骨再生是重要的。目标3将验证ARS中的最佳扩张过程，并在异位小鼠模型中评估体内骨和血管再生。目标4将包括一项I/II期临床试验，使用选定的细胞产品与基质材料组合治疗胫骨骨不连骨折。这些研究的成功完成将导致可用于骨再生的细胞治疗试剂盒的开发。

DESCRIPTION (provided by applicant): Due to morbidity and limited amounts of material, alternatives to autologous grafts for bone repair are desirable. Cells within the bone marrow are known to possess bone regenerative capacity, but they are limited in number. Increasing these osteoprogenitor cell populations through the use of ex vivo expansion technologies and combining these cells with a matrix material may significantly increase the amount and rate of new bone formation, and replace the need for autologous grafts. Phase I studies identified expansion variables for osteoprogenitor cell expansion and showed that this process can be scaled up into the automated clinical-scale cell culture system, the AastromReplicell(r) Cell Production System (ARS). In Aim 1 of the Phase II studies, optimization of the osteoprogenitor expansion process will continue. Optimization will be measured by not only biological performance in vitro and in vivo, but also the ease of commercialization of the expansion process. Specifically, the role of other cell populations in osteoprogenitor cell expansion and function will be determined, the minimal requirements for including exogenous factors in cultures will be identified, inoculation densities and maximized feeding schedules will be determined to support expansion from small volume bone marrow aspirates, the potential for expansion in an animal serum-free environment will be examined. In Aim 2, the angiogenic potential of these same cell products will be measured in vitro by colony assay and tube formation assays, as blood vessel formation is important for optimal bone regeneration. Aim 3 will validate optimal expansion processes in the ARS and evaluate bone and vascular regeneration in vivo in an ectopic mouse model. Aim 4 will include a Phase l/ll clinical trial using the selected cell product in combination with matrix material to treat tibial non-union fractures. Successful completion of these studies will lead to the development of a cell therapy kit that can be used for bone regeneration.